GENA PHARMACEUTICALS LIMITED

KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 1 of 13

CLEANING VALIDATION
PROTOCOL CUM REPORT
FOR
SAMPLING ACCESSORIES
 GENA PHARMACEUTICALS LIMITED
KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 2 of 13

TABLE OF CONTENTS

S. No. TITLE PAGE NO.

1. Protocol pre-approval 1

2. Objective 2

3. Scope 2

4. Reference 2

5. Site of Study 2

6. Responsibility 2

7. Materials and instruments 3

8. Cleaning Procedure 3

8. Experimental Design 3

9. Acceptance Criteria 4

10. Result 4

11. Conclusion 4

12. Validation Methodology 4

13. Calculation 5

14. Cleaning Validation Datasheet 7

15. Determination Of Traces of Alpha Lipoic Acid 7

16. Determination Of Traces of Teepol 10

17. Cleaning Validation Report 11

 GENA PHARMACEUTICALS LIMITED
KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 1 of 13

1.0 PROTOCOL PRE- APPROVAL

INITIATED BY:

S. No NAME DESIGNATION SIGNATURE DATE

1.

REVIEWED BY:

S. No NAME DESIGNATION SIGNATURE DATE

2. Executive QA

3. QC Manager

APPROVED & AUTHORISED BY:

S. No NAME DESIGNATION SIGNATURE DATE

4. Head- QA
 GENA PHARMACEUTICALS LIMITED
KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 2 of 13

1.0 OBJECTIVE: To provide the documentary evidence to ensure that the procedure used for the
cleaning of sampling accessories is effective.

2.0 SCOPE:

This validation study is performed on the sampling accessories which are used during routine
sampling of raw material.

3.0 REFERENCE:

SOP NO:- QC/014-01 :-Coding and cleaning of sampling device.

4.0 SITE OF STUDY:

Quality Control Laboratory

5.0 RESPONSIBILITY:

DEPARTMENT RESPONSIBILITIES

Review, Approval and Compilation of the Cleaning Validation Protocol for

QUALITY Correctness, Completeness and Technical Excellence.
ASSURANCE
Monitoring of validation Activity.

Co-ordination with Quality assurance to execute the protocols.

QUALITY Preparation ,Review of Cleaning validation Protocol, testing and
CONTROL
Execution.
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KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 3 of 13

DEFINITION
Sampling Accessories:- The accessories which are used to withdraw the material e.g spatula, scoop,
liquid sampling device, solid sampling device ,chopper

6.0 MATERIALS AND INSTRUMENTS:

a) Instruments : UV Visible Spectrophotometer and HPLC
b) Cleaning agent : Teepol

7.0 CLEANING AND STORAGE PROCEDURE:

7.1 Cleaning of sampling accessories as follows:-

7.2 Wash all the sampling accessories in tap water.
7.3 Rub the inner and outer surface of of sampling device using liquid soap ( 2%Teepol) with plastic
brush
7.4 Rinse the sampling accessories in running tap water.
7.5 Wipe inner and outer surface of sampling device using lint free cloth moist in 70% isopropyl
alcohol
7.6 Affix a label on polyethylene bag containing sampling device.
7.7 The sampling device used for microbial testing to be cleaned as per above procedure and then
sterilize at 1210C, 15lbs for 30 minute

TYPES OF SAMPLING ACCESSORIES

SAMPLING ACEESORIES USED FOR SAMPLING

8.0 EXPERIMENTAL DESIGN:

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KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 4 of 13

1) Select the labeled sampling device as mentioned for the sampling of Alpha Lipoic Acid.
2) Isolate the sampling device in sealed polythene bag immediately after sampling.
3) Monitor the movement of the selected sampling device during washing, take care to maintain its
identity throughout the cleaning.
4) Clean the respective sampling device as per routine cleaning procedure mentioned in SOP and dry
the same.
5) Analyze the rinse sample as per mentioned in method details and report the observations in the
cleaning validation data sheet.
6) Rinse the volumetric flask with 10 ml of water and determine traces of Teepol.

9.0 ACCEPTANCE CRITERIA:

Finding of the study should be below detectable limit as specified in the method details.

10.0 RESULT:
1) Observation and results to be recorded in the respective cleaning validation data sheet.
2) Summarize the finding of the study to draw inference.

11.0 CONCLUSION:
Based on the interpretation of the results in cleaning validation summary, draw the conclusion for
the effectiveness of the cleaning procedure used.
12.0 VALIDATION METHODOLOGY:
A) Determination of traces of Alpha Lipoic Acid
Chromatographic Conditions :
Column :A Stainless steel column 250mm X 4.6mm(5µ),C18
Flow rate :1.2 ml/min
Detector :215nm
Injection Volume :20μl
 GENA PHARMACEUTICALS LIMITED
KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 5 of 13

Diluent :Mobile phase

Blank Injection :1 run(Diluent)
Standard injection :5 run
Sample injection :2 run

Preparation of mobile phase:

Buffer Preparation: Dissolve 340 mg Potassium dihydrogen phosphate in 500ml Distilled water.
Preparation of Mobile Phase: Buffer: Methanol: Acetonitrile=230:290:45
Adjust the PH 3.0 to 3.1

Preparation of Standard solution:

Weigh and transfer accurately about 25mg Alpha Lipoic Acid WS into 25ml volumetric flask. Add 10
ml of diluent and sonicate for 10 minutes. Make up the volume upto 25ml with mobile phase.Further
dilute 1ml of this solution to 100 ml with mobile phase.
Preparation of Sample solution:
Add 10ml of diluent with the help of pipette in the cleaned accessories. Swirl the device determine
content . Determine the content in rinse sample collected.

13.0 CALCULATION:

Average area of sample X Standard wt X 1 X Purity X1000

Average area of standard X25 X 100 X 100

Limit of Detection
Alpha Lipoic Acid = 0.1ppm

B) Determination of Traces of cleaning agent (Teepol):

 GENA PHARMACEUTICALS LIMITED
KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 6 of 13

Preparation Of Standard (Teepol):

Weigh 100mg of Teepol in a 100ml volumetric flask, dissolve and dilute to the volume with water.
Dilute 10ml of this solution to 100ml with water. Further dilute 7.0 ml of this solution to 50 ml with
water.
Sample Preparation:
Add 10 ml of water with the help of pipette in the cleaned sampling device. Swirl the device and
determine content in rinse sample collected.

Calculation:
Measure the absorbance of Teepol solution at 223 nm of the sample solution using water as blank.

Traces of Teepol (ppm):

= Sample absorbance X Standard weight (mg) X 10 X 7.0 X 1000

Standard absorbance X 100 X 100 X 50

Limit of Detection: 2.0ppm

CLEANING VALIDATION DATA SHEET

 GENA PHARMACEUTICALS LIMITED
KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 7 of 13

A) Determination Of Traces of Alpha Lipoic Acid

Date:- Inst. ID:
Sign: Make:
Balance ID: Model No:

Chromatographic Condition:
Column :A Stainless steel column 250mm X 4.6mm(5µ),C18
Flow rate :…… ml/min
Detector :……nm
Injection Volume :……μl
Diluent :…………….
Blank Injection :1 run (Diluent)
Standard injection :5 run
Sample injection :2 run
Chemicals Used:
1. Potassium Dihydrogen phosphate B.No./Lot.No:
2. Methanol B.No./Lot.No:
3. Acetonitrile B.No./Lot.No:

Preparation of mobile phase:

Preparation of buffer solution:
Monobasic potassium phosphate taken = …….. g (~0.68 g).
Dilution = ……… ml (1000 ml) with distilled water.

Preparation of mobile phase: Methanol: Buffer: Acetonitrile = 230:290:45

pH adjusted = …… (3.0 to 3.1).

Preparation of Standard solution:

 GENA PHARMACEUTICALS LIMITED
KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 8 of 13

Standard taken = …… mg (~25 mg).

Standard dilution = …… ml (25 ml).
= … ml (1 ml) of resultant solution in diluted to …. ml (100 ml) with mobile phase.

Preparation of Sample solution:

Add ……..ml of diluent with the help of pipette in the cleaned sampling device. Swirl the device and
determine content in rinse sample collected.
Standard Used:
Alpha Lipoic Acid WS ID No:
% Purity:
Average Standard area=(…………….+…………….+……………….+……………+………………)/5
=………………..
Sample Area
After 1st washing After 2nd washing After 3rd washing
Device Area Average Device Area Average Device Area Average
used area used area used area

Calculation: Calculate the traces of Alpha Lipoic Acid as per formula mentioned below.
Traces of Alpha Lipoic Acid (ppm):
 GENA PHARMACEUTICALS LIMITED
KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 9 of 13

= Average area of sample X Standard wt X 1 X Purity X1000

Average area of standard X25 X 100 X100

Limit of detection:-0.15ppm
Stages of Rinsing
After 1st Rinsing After 2nd Rinsing After 3rd Rinsing
Device used Avg. Result Device used Avg. Result Device used Avg. Result
Area (ppm) Area (ppm) Area (ppm)

CLEANING VALIDATION DATA SHEET

A) Determination Of Traces of Teepol
Date:- Inst. No:
 GENA PHARMACEUTICALS LIMITED
KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 10 of 13

Sign: Inst.Make:
Balance ID: Inst.Model:
Standard Used:
Teepol WS ID No:
% Purity:
Standard Absorbance:

Calculation:
Measure the absorbance of Teepol solution at 223 nm of the sample solution using water as blank and
calculate as per mentioned below .
Traces of Teepol(ppm):
= Sample absorbance X Standard weight (mg) X10 X 7.0 X 1000
Standard absorbance X 100 X 100 X 50
Limit of detection:-2ppm
Stage of Rinsing
After 1st Rinsing After 2nd Rinsing After 3rd Rinsing
Sampling Abs. Result Sampling Abs. Result Sampling Abs. Result
device used (ppm) device used (ppm) device used (ppm)

CLEANING VALIDATION REPORT

RESULTS:
 GENA PHARMACEUTICALS LIMITED
KOLKATA - 700128
CLEANING VALIDATION OF SAMPLING ACCESSORIES
Protocol No. Revision No. 00 Page No.:

Effective Date Review Date: 11 of 13

Sampling device used Content of Alpha Lipoic Acid Limit of Detection

Sampling device used Content of Teepol Limit of Detection

CONCLUSION: The above result shows that traces of previous drug substances and cleaning agent
used for cleaning is present with in accepted limit and hence the procedure used for the cleaning of
sampling accessories stands validated.